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. 2019 Aug 27;2019:10.17912/micropub.biology.000146. doi: 10.17912/micropub.biology.000146
Plasmids
pALS297 (ttTi5605-MCS loxP::Prps-27::NeoR::unc-54-3ʹutr::loxP); Addgene # 129538
pALS186 (ttTi5605-MCS loxP::C.br.unc-119(+)::loxP); Addgene # 129539
Strains
EG6699 ttTi5605(mos1) II; unc-119(ed3), CGC, note lost extrachromosomal array oxEx1578
ALS344 ttTi5605(mos1) II, this study; EG6699 was crossed 4x to N2 to remove the unc-119(ed3) allele
Primers to follow ttTi5605(mos1) II
o508-mos1F(oJL103) TCTGCGAGTTGTTTTTGCGTTTGAG
o509-ttTi5605F GATTGTTTGACCTGGCGGAACT
o91-ttTi55605R(NM3888) ACGCCCAGGAGAACACGTTAG
Primers for cloning
op297-p186AvrIIF TAGAGGGTACCAGAGCTCACCTAGGN*20-22
op297XhoIR GTTATaCGCGCACCGTACGTCTCGAGN*20-22
op186XhoIR GTTATgCGCGCACCGTACGTCTCGAGN*20-22
Primers for insert verification
5ʹarm, neoR mosSCI
o145-ttTi5605outerF AGGCAGAATGTGAACAAGACTCGAG (NM3880 + 2nt)
o496-unc54-3ʹutrR GGCCCAGACGTGCGAAGAAATA

(or transgene-specific REV primer for long transgenes)

3ʹarm, unc-119 mosSCI
o145-ttTi5605outerF AGGCAGAATGTGAACAAGACTCGAG (NM3880 + 2nt)
o147-Cbr-unc-119promR AAGTAGCAGAGCTGGGGAGAAGAA

(or transgene-specific REV primer for long transgenes)

5ʹarm, neoR mosSCI
o311-NeoRF CTTCTATCGCCTTCTTGACGAG
o146-ttTi5605outerR AATCGGGAGGCGAACCTAACTGT (NM3884 + 2nt)
3ʹarm, unc-119 mosSCI
Transgene-specific FOR primer
o146-ttTi5605outerR AATCGGGAGGCGAACCTAACTGT (NM3884 + 2nt)